This study examined how the addition of a sawdust extract from Castanopsis cuspidata, several aromatic compounds, and copper affected the expression of a phenol-oxidizing enzyme in the white-rot basidiomycete, Lentinula edodes. Compared to liquid media that had not been supplemented with sawdust extract (MYPG), MYPG containing low (MYPG-S-100) or high (MYPG-S-500) concentrations of sawdust extract had a marked effect on the promotion of mycelial growth. No manganese peroxidase (MnP) production was observed in either MYPG or MYPG-S-100 media until 35 days after inoculation. However, MnP production was enhanced by culture in MYPG-S-500, with a marked increase observed suddenly at 14 days after inoculation. Northern blot analysis revealed that the transcription of the lemnp2 gene coding extracellular MnP was initially observed at detectable levels at day 10 after the initial inoculation of MYPG-S-500, increasing gradually thereafter until days 22-25. However, laccase (Lcc) production was not observed in any of the media until 35 days after inoculation. Addition of 10 mM aromatic compounds - 1,2-benzenediol, 2-methoxyphenol, hydroquinone, and 4-anisidine - into the MYPG-S-500 medium completely inhibited MnP production and did not enhance any Lcc production. While the addition of 1 or 2 mM Cu2+ (CuSO4 center dot 5H(2)O) to MYPG-S-500 medium completely inhibited MnP production, this Cu2+ addition caused a marked increase in Lcc production at 17 and 6 days after the addition, respectively.

Sesamum mulayanum is a wild relative of cultivated sesame, Sesamum indicum, and sometimes grows in sesame crop fields as an associated weed. This species shows deep seed dormancy and is characterized by conspicuous purple pigmentation on the lower lip of the corolla. The present study examined the inheritance mode of seed dormancy by using reciprocal progeny from crosses between the two species. The seeds of S. indicum and F1 (S. indicumXS. mulayanum) showed good germination, but those of S. mulayanum and F1 (S. mulayanumXS. indicum) showed deep dormancy. The F2 seeds from both reciprocal crosses showed deep dormancy. These results, combined with the maternal inheritance of seed-coat characteristics, indicated that the seed dormancy of S. mulayanum can be attributed to its seed-coat structure (coat-enhanced dormancy). The F3 (S. indicumxS. mulayanum) seeds varied in their depth of seed dormancy and those seeds with deep dormancy (<50% germination) and those with no or shallow dormancy (=50% germination) occurred in the expected ratio of 3:1, indicating that this trait is polygenic but is controlled by a single dominant major gene. The purple pigmentation of the corolla was expressed in both reciprocal F1 plants and the presence and absence of pigmentation was segregated among the F2 plants at the expected ratio of 3:1, indicating that this trait is also controlled by a single dominant gene. The segregation of the major gene controlling seed dormancy and that controlling purple pigmentation was not independent (9:3:3:1), indicating that these genes are linked, providing insights on sesame domestication.

Electrophoretic karyotype analyses of Flammulina velutipes FSB and its monokaryotic progeny, omFSB1 and omFSB2, obtained from oidia were performed by contour-clamped homogeneous electric field (CHEF) gel electrophoresis. At least 11 chromosome-sized DNA bands (CB 1 through CB 11) for FSB, 6 bands for omFSB1, and 7 bands for omFSB2, respectively, were resolved on a CHEF gel. Southern hybridization analysis on CHEF-separated chromosomal DNA of FSB was carried out using RAPD marker probes prepared from each of the 12 linkage groups. The bands CB 1, 2, and 4 each hybridized to two or three probes for different linkage groups. The bands CB 5 and 6 both hybridized to a common probe. The bands CB 3, 7, 8, and 9 each hybridized to a single specific probe for different linkage groups. The two smallest bands (CB 10 and 11) did not hybridize with any probes.

The biocliversity of saprotrophic and ectomycorrhizal basidiomycetous macrofungi growing on seven islands in central japan were compared to examine colonizing success within the context of island biogeography theory. Two hypotheses were tested: that the number of the fungal species depends on island area and that the slope of the species-area curve for saprotrophic and ectomycorrhizal macrofungi differ in response to differences in their nutritional requirements. Data for the number of species that were identified based on sporocarps closely fit the conventional species-area curve. The slopes of the species-area curve for saprotrophic fungi (0.316) and ectomycorrhizal fungi (0.469) were similar to those reported for insects and birds, and plants on other archipelagos, respectively. In addition species-area curve data showed that ectomycorrhizal fungi colonized only islands > 630 m(2). While the species composition of saprotrophic fungi found on any pair of islands was positively correlated to the ratio of the areas of the island pair being compared (smaller/larger), no such relationship was observed for ectomycorrhizal fungi. Conversely similar ectomycorrhizal fungi, mostly those belonging to the genera Amanita, Inocybe, Boletellus and Russula, were found on pairs of islands with similar vegetation in the same geographic region. These results suggested that the colonizing success by ectomycorrhizal fungi is limited by host plant diversity, which is lower on smaller islands, instead of restricted immigration resulting from limited spore dispersal ability.

We tried to produce new varieties of sweet potato, Ipomoea batatas (L.) Lam. which are superior in taste and rich in functional pigments. This was carried out through cell fusion between sweet potato cultivars, Ariakeimo, Purple sweet lord and Benihayato. First, the density of protoplasts at the fusion stage and the composition of initial culture medium were investigated. As a result, through the fusion between Ariakeimo and Purple sweet lord at a density of 1.0×10⁵ protoplasts/ml in dark at 25℃, the visible micro-callus of about 2〜3 mm in diameter, was formed on N6 medium supplemented with 0.1 mg/l 2,4-dichlorophenoxyacetic acid, 0.5 mg/l kinetin, 0.2 M sucrose and 0.6 M mannitol, pH5.8. Second, the composition of efficient regeneration medium was investigated in the plant regeneration from petiole callus of Ariakeimo, Purple sweet lord and Benihayato. Eventually it became obvious that individual shoot induced from the callus have grown into young plant at a high rate on the MS medium supplemented with 0.03 mg/l α-naphthaleneacetic acid, 0.01〜0.09 mg/l 6-benzyladenine, 0.087 M sucrose and 1.2% agar.

Seed germination of sesame (Sesamum indicum L.) and its wild relative S. mulayanum Nair was examined, focusing on dormancy break of S. mulayanum. S. indicum showed higher germination (%) at 30 and 35℃ as compared to that at 25℃, while S. mulayanum showed deep dormancy and failed to germinate at all temperatures tested. Soaking seeds in gibberellic acid (GA3) solution had no effect on seed germination of S. mulayanum at any concentration (5, 10, 50, 100 and 200μM) of GA3 tested. Treatment of the scarification promoted seed germination of both S. indicum and S. mulayanum and completely broke seed dormancy of S. mulayanum resulting in 90% germination. The present method for seed dormancy break is expected to aid the examination of inheritance mode of seed dormancy in the crossed progeny between the two species.

The white-rot basidomycete Lentinula edodes often produces the lignin-degrading enzymes manganese peroxidase (MnP; EC 1.11.1.13) and laccase (Lcc; EC 1.10.3.2) in sawdust-based media. In the present study, MnP from L. edodes was induced under liquid culture supplemented with sawdust extracts of Castanopsis cuspidata. Lcc activity was induced by the addition of 2 mM CuSO4 center dot 5H(2)O into the same media 7 days after initial inoculation. Phenoloxidase enzymes were distinguished by polyacrylamide gel electrophoresis (native-PAGE), followed by sequential enzymatic staining with an improved staining solution. The isozyme bands detected under MnP-induced conditions were identified as manganese peroxidase (lemnp2) and bands detected under Lcc-induced conditions were identified as laccase (lcc1) by Q-TOF mass spectrometry.

The natural habitat of the Echinochloa species growing in eastern Australia was investigated in May 2008. In this survey, four annual native species, E. dietrichiana, E. inundata, E. telmatophila, and E. turneriana, and three exotic species, E. colona, E. crus-galli, and E. polystachya, were observed. The native species clearly have separate habitats: permanent stagnant pools for E. dietrichiana, inland floodplains for E. inundata, in or beside streams for E. telmatophila, and inland watercourses for E. turneriana. While the habitat of the exotic species greatly overlap that of the natives, especially perennial E. polystachya, which forms monospecific stands in stagnant pools and in or beside flood rivers due to its remarkable productivity, and E. crus-galli, which has the potential to adapt to several moisture conditions. We predict that the invasive exotic Echinochloa species will become more and more threatening causing serious damage to the native species within the same genus.

紀伊半島南西海岸沖の 4 つの島において、 生息するきのこ類を調べた。 数種の小型腐生菌はどの島でも観察される定着種として挙げられた。 生息種数の対数は島面積の対数に比例したが、 樹木と共生する外生菌根菌の定着率は極めて低かった。

奈良県平群町において約 200 種のきのこ類を同定した。 また、 平群町住民によって伝統的に利用されるきのこ類がアカマツ林に依存する数種に限られることを示した。

林床のリター分解に関わる菌体外酵素の活性を測定し､ リター成分中のリグニンとセルラーゼの分解に関わる酵素群の活性について､ 季節推移のパターンを示した｡ さらに F 層における各種酵素活性の高さから､ 本法による微生物生理活性の推定の妥当性を示した｡

菌床培地において、正常株nは全ての栽培瓶で子実体を形成した。変異株d3とd4は一部の栽培瓶で菌床表面の一部のみから子実体を形成したことから、両株は子実体形成に関して極めて不安定であるものの、完全な子実体形成不能ではないと判断した。一方、変異株d5は菌床面に子実体原基の様な凹凸は観察されたが、それ以上の分化はみられなかったことから、完全な子実体形成不能株であると判断した。フラスコ内のMYS液体培地において、正常株nでは25℃で培養した栄養菌糸体を15℃の低温下に培養温度を変更することで子実体を形成したが、変異株d3、d4、d5では低温下でも子実体は形成されなかった。 RNA-seq解析において、エノキタケの子実体形成期に特異的に発現することが知られているFDS遺伝子やエノキタケを含む数種の菌類でも子実体形成期に発現するhyd1遺伝子が、低温刺激を与えた正常株でのみ誘導される遺伝子としてピックアップされた。RT-qPCRによる子実体形成関連遺伝子の発現解析において、Fvhyd1遺伝子とFvFDS1遺伝子は低温刺激を与えた、変異株d3、d4では発現せず、正常株nの原基形成期に特異的に発現していた。しかし、低温刺激を与えた、変異株d5では両遺伝子の発現が確認されたことから、この2つの遺伝子は子実体原基形成において、必要条件であるが十分条件では無いと考えられた。また、両遺伝子の発現はすべての菌株で連動していたことから、これらの発現を制御する共通の転写調節因子の存在が考えられる。本研究によって、これまで野生株を用いた子実体形成期に発現すると観察されていたFDS1やhyd1遺伝子の特性が、子実体形成不全株を用いることで、より明確に示された。

Compatibility of three bacterial strains in Agrobacterium tumefaciens-mediated transformation (AMT) of haploid-fruiting strain (bmHY4) of Flammulina velutipes was compared. The bacterial strains LBA4404 and C58C1 yielded four- to five-fold higher transforming efficiencies than did EHA105. Nevertheless, the transforming efficiencies observed in this study are still not sufficient for practical construction of a F. velutipes mutant library. Then we attempted to obtain transposon tagging mutants. An autonomously transposable element impala from a fungus Fusarium oxysporum was ligated into a binary vector pPZP-Hyg2 and successfully transformed bmHY4 mycelia via AMT.

Biodiversity of genus Echinochloa (Poaceae) was surveyed by field expedition in several target areas of the Eurasia, Africa, North and South America, and Oceania regions, and by herbarium collections at K, CAL, UC, KUN etc. Annual species, both native and alien, have potential risks as rural weeds including herbicide resistant biotypes, and perennial species mainly alien as invaders in natural aquatic habitats.

It has been shown that carbohydrate molecules play an important role in multicellular formation : fertilization, development, differentiation, immunity, and canceration. It is quite difficult to show the evidence concerning the function of carbohydrate molecules, due to the weakness of binding abilities between carbohydrate molecules and the target molecules. In this study, our focus was on analyzing the carbohydrate function in multicellular formation through cell adhesion glycoproteins, intrinsic lectins, the extracellular, matrix, and so on. Moreover our aim was to clarify the function of carbohydrate molecules, using mutants defective in carbohydrate molecules and carbohydrate genes. We tied to isolate various kinds of carbohydrate mutants, taking advantage of the special properties of Dictyostelium discoideum as a haploid organism. After Dictyostelium wild-type cells were treated with mutagens, cells were allowed to grow for 4 to 5 days. Negative cell groups were isolated and pooled as the first screening by using FTT-conjugated lectins: Con A, WGA, PNA, MAL and UEAI and a flow cytometer, FACS Vantage SE. Next, among negative cell groups in the first screening, each negative cell was isolated by a flow cytometer in the second screening. As a result, several mutants were isolated by using C-conjugated Con A Western blotting analysis showed several changed bands with Con A reactivities in the mutant cells. The growth rate of the mutant cells was much slower than that of wild-type cells.